农吉利碱对乳腺癌细胞MDA-MB-231凋亡及相关因子表达的影响

Influence of monoerotaline on apoptosis and related factors of MDA-MB-231 cells

目的:探讨农吉利碱对乳腺癌细胞MDA-MB-231增殖与凋亡的影响。方法:体外培养MDA-MB-231细胞,给予不同药物浓度(0、10、20、40μmol/L)农吉利碱处理MDA-MB-231细胞24、48、72h。四甲基偶氮唑盐比色(MTT)法检测对MDA-MB-231细胞活力的影响,光镜和吖啶橙染色分析细胞形态,流式细胞术检测农吉利碱处理细胞72h后的凋亡率和细胞内活性氧水平,免疫印迹法(Western blot)检测B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、激活型半胱氨酸蛋白酶-3(Cleaved-Caspase-3)蛋白表达。结果:农吉利碱10、20、40μmol/L对MDA-MB-231细胞具有明显抑制作用,光镜、吖啶橙染色法可见细胞出现凋亡特征性改变,农吉利碱可诱导细胞凋亡,抑制Bcl-2蛋白表达,促进Bax和Cleaved-Caspase-3的蛋白表达,并与农吉利碱浓度呈依赖性。结论:农吉利碱抑制乳腺癌细胞MDA-MB-231增殖,并促进其凋亡,与Bax和Cleaved-Caspase-3的表达增加和Bcl-2降低有关。

Objective： To explore the effect and mechanism of moncorotaline on apoptosis of breast cancer ceils MDA-MB-231 in vitro. Methods： Breast cancer cell MDA-MB-231 cells were cultured, and treated by moncorotaline at concentrations of 0,10,20,40 μmol/L and at 24,48 and 72h. Effect of moncorotaline on breast cancer cells activity was detected by 3- （4,5-dimethyl-2-thiazolyl） -2,5-diphenyl-2-H-tetrazolium bromide （MTr） assay; cells morphology was observed by phase contrast microscope, cell apoptosis and intracellular levels of reactive oxygen species were measured by flow cytometry after treated by moncorotaline for 72h. Protein expressions of Bcl-2 and Bax, Cleaved-caspase-3 were detected by Western blot. Results： Monocrotaline （10,20,40pJnol/L） could significantly inhibit the viability of MDA-MB-231 cells, typical apoptosis changes of cells were showed by light microscope and acridine orange staining methods ; thus monocrotaline could induce ceil apoptosis, inhibit the protein expression of Bcl-2 and enhance the protein expression of Bax and Cleaved-Caspase-3 in a concentration dependent manner. Conclusion： Moncorotaline significantly inhibited the activity of breast cancer cells, and induced apoptosis in concentration-depend- ent manners. Protein expressions of Bax and Cleaved-Caspase-3 were up-regulated by moncorotaline, while Bcl-2 expression was inhibited,